KR20130010096A - Kit for skin beauty including cosmetic composition and method for applying the cosmetic composition - Google Patents

Abstract

PURPOSE: A skin care kit and a coating method are provided to show an excellent skin cooling, skin elasticity enhancement and skin wrinkle diminishing effect by coating a cosmetic composition having the skin elasticity enhancement effect using a skin application apparatus providing the tensile stress. CONSTITUTION: A skin care kit comprises a cosmetic composition containing arginine and lysine-containing polypeptide as active ingredients and a skin application apparatus for coating the cosmetic composition on the skin. The skin application apparatus is at least one selected from the group consisting of a roller, a pressure indicator and massage device providing the tensile stress. The coating of the cosmetic composition comprises a step of applying the tensile stress to the skin using the skin application apparatus while or after coating the cosmetic composition on the skin. [Reference numerals] (AA) Fibroblast under tensile stress; (BB) Widening and reinforcing actin; (CC) Before stimulation; (DD) After stimulation

Description

KIT FOR SKIN BEAUTY INCLUDING COSMETIC COMPOSITION AND METHOD FOR APPLYING THE COSMETIC COMPOSITION}

The present invention relates to a skin care kit and a coating method comprising a cosmetic composition, and more particularly to a skin care kit and a coating method comprising a cosmetic composition excellent in the skin cooling effect by tensile stimulation.

In general, cosmetics are intended to clean the body, to make themselves beautiful and attractive, and to prevent aging by protecting the skin or hair from ultraviolet rays or drying. Recently, with the development of industrial society and the increase of social activities, the self-consciousness to actively express oneself has increased, and the desire to care for and maintain one's own skin by using cosmetics has greatly increased. Cosmetics are being researched and developed.

Skin elasticity is represented by elastic fibers composed of elastin in the dermis, which has a very low modulus of elasticity, such as rubber, which is easily deformed by small forces, and easily removed when the force is removed. Come back to In addition, elastic fibers have a form in which microfibrils are embedded in an amorphous substrate called elastin, and elastin is a very unique feature found only in elastic fibers called desmosine and isodesmosine derived from lysine. It is a protein consisting of one amino acid. These desmosin and isodesmosin form cross-links in long peptide chains, which give elastin a rubbery property.

In addition, the elastic fiber composed of elastin is present together with collagen fibers called collagen, and it is possible to maintain skin elasticity in a state where elastin and collagen are sufficiently present.

On the other hand, the decrease in skin elasticity is caused by a decrease in the production and destruction of elastin due to the weakening of skin cells (fibroblasts) due to aging and ultraviolet light. Elastin degrading enzyme is expressed in the skin by ultraviolet rays, and this enzyme decomposes normal elastin in the skin, resulting in a decrease in skin elasticity. In addition, a decrease in skin elasticity is also caused by damage to and production of a collagen protein, which makes up more than 70% of the skin dermis. Collagen is also broken down by a degrading enzyme called collagenase, which is caused by ultraviolet light. Existing patents related to the enhancement of skin elasticity simply inhibit the activity of collagenase or suppress the activity of elastase in order to show the skin elasticity enhancing effect, has a limited effect on the dermis layer to maximize the function There was a disadvantage that can not come.

On the other hand, as the polarization of the climate has progressed in recent years, there is a growing interest in products that can lower and cool down the skin temperature in the summer as the summer becomes longer and hotter. In fact, the makeup is clogging the pores of the skin and gives a hot feeling, and when the sweat flows out, the need for cool cosmetics is increasing among women who need daily makeup. Accordingly, as summer products, products are aimed to cool down skin that has been heated by heat to cool down, tighten pores, and improve mood. Specifically, there are many products that use ingredients such as menthol, menthol derivatives, xylitol, and the like as cooling agents in general cosmetic formulations, but it is not enough to feel an immediate cooling effect.

According to the present invention, a cosmetic composition containing arginine and a lysine-containing polypeptide as an active ingredient is applied to the skin using a skin applicator, thereby exhibiting excellent skin cooling effect by tensile stimulation that cannot be realized by a general application method. It is an object to provide a kit and a coating method.

In order to solve the above object, the present invention provides a skin care kit comprising a cosmetic composition containing arginine and lysine-containing polypeptide as an active ingredient, and a skin coating device for applying the cosmetic composition to the skin.

In one embodiment of the present invention, the skin applicator may be one or more selected from the group consisting of a roller, acupressure and a massage device to give a tensile stimulus.

In one embodiment of the present invention, the skin applicator may be a roll-on (roll-on).

In one embodiment of the present invention, the roll-on may be three or more balls.

In one embodiment of the present invention, the cosmetic composition may be one that further contains one or more of oat extract and yeast extract.

In one embodiment of the present invention, the cosmetic composition is contained in 0.01 to 2% by weight of the arginine and lysine-containing polypeptide, 2 to 6% by weight oat extract and 0.01 to 0.5% by weight yeast extract relative to the total weight of the composition Can be.

In one embodiment of the present invention, the cosmetic composition may be one that further contains one or more selected from the group consisting of adenosine, retinol and niacinamide.

In one embodiment of the present invention, the skin care kit may be one having at least one selected from the group consisting of skin cooling use, skin elasticity promotion use and skin wrinkle improvement use.

In addition, the present invention provides a cosmetic composition comprising the step of applying a tensile composition to the skin using a skin coating device after or simultaneously with the application of the cosmetic composition containing the arginine and lysine-containing polypeptide as an active ingredient to the skin, Provide a coating method.

In one embodiment of the present invention, the coating method may be one having at least one selected from the group consisting of skin cooling use, skin elasticity enhancement use and skin wrinkle improvement use.

According to the present invention, by applying a cosmetic composition having a skin elasticity enhancing effect to the skin using a skin coating device that gives a tensile stimulus rather than a general coating method, the synergistic effect of both the cosmetic composition and the coating method occurs to produce more excellent skin It can provide a skin care kit and application method that exhibits cooling, skin elasticity enhancement and skin wrinkle reduction effect.

1 is a photograph showing a change in actin during tensile stimulation according to an embodiment of the present invention.
Figure 2 is a graph of the relative expression of collagen when using a cosmetic composition and tensile stimulation according to an embodiment of the present invention.
3 is a cross-sectional view of a skin coating device according to an embodiment of the present invention.
Figure 4 is a perspective view of the upper ball installation portion (a) and the lower ball installation portion (b) of the skin coating apparatus according to an embodiment of the present invention.
5 is a graph showing the measurement results of the skin cooling experiment when using the cosmetic composition and the skin coating apparatus according to an embodiment of the present invention.
Figure 6 is a graph showing the skin temperature decrease rate when using the cosmetic composition and the skin coating apparatus according to an embodiment of the present invention.
Figure 7 is a photograph of the eye skin temperature when using the cosmetic composition and the skin coating apparatus according to an embodiment of the present invention.
Figure 8 is a graph showing the measurement results of the eye area wrinkles test when using the cosmetic composition and the skin coating apparatus according to an embodiment of the present invention.
Figure 9 is a graph showing the wrinkle reduction of the eye area when using the cosmetic composition and the skin coating apparatus according to an embodiment of the present invention.
10 is a photograph of the wrinkles around the eyes when using the cosmetic composition and the skin coating apparatus according to an embodiment of the present invention.
Figure 11 is a graph showing the eye area elasticity increase rate when using the cosmetic composition and the skin coating apparatus according to an embodiment of the present invention.
12 is a graph showing the measurement results of the eye area elasticity test when using the cosmetic composition and the skin coating device according to an embodiment of the present invention.
13 is a graph (a) and blood flow image (b) showing skin blood flow when a cosmetic composition and a skin coating apparatus according to an embodiment of the present invention are used.
14 is a graph (a) and absorption rate (b) of the absorption amount of the active ingredient when using the cosmetic composition and the skin coating apparatus according to the embodiment of the present invention.

Hereinafter, preferred embodiments of the present invention will be described in detail in order to facilitate the present invention by those skilled in the art.

The present invention provides a skin care kit comprising a cosmetic composition containing arginine and a lysine-containing polypeptide as an active ingredient, and a skin applicator for applying the cosmetic composition to the skin. The cosmetic composition itself has the effect of cooling the skin, enhancing skin elasticity and improving skin wrinkles.

When the cosmetic composition described above is applied to the skin, it is applied while giving a tensile stimulus to the skin using a skin coating device. The skin applicator is not particularly limited, but may be one or more selected from the group consisting of a roller, acupressure and a massage device that give a tensile stimulus.

Tensile stimulation is a physical stimulus that expands and contracts the skin by applying pressure to the skin. Tension stimulation strengthens the cytoskeletal proteins of skin cells. As can be seen in Figure 1, actin in fibroblast cytoskeletal protein is strengthened during tensile stimulation.

In addition, tensile stimulation expresses anti-aging and barrier function improvement genes of skin cells. Related research results are as follows.

In addition, tensile stimulation improves the blood flow of the skin and promotes skin absorption of the substance. It also helps the skin's immediate cooling, wrinkle and elasticity. This will be confirmed in the following experimental example.

Tensile stimulation to the skin using the skin coating device generates a massage effect and the effect of promoting the penetration of the active ingredient, so that the skin elasticity enhancing effect of the cosmetic composition containing the arginine and lysine-containing polypeptide as an active ingredient can be more exerted. help. In addition, the skin coating device is in direct contact with the skin to conduct the heat of the skin and apply a tensile stimulation can be exerted more excellent skin cooling effect. That is, synergistic effects of skin cooling, skin elasticity enhancement and skin wrinkle improvement are generated by the simultaneous use of the cosmetic composition and skin applicator.

Preferably the skin coating device may be a roll-on (Roll-on). The roll-on may have one, two or three or more balls. Such roll-on is not limited as long as it makes the ball | bowl of a spherical form contact skin.

In one example, a roll-on having three balls in the skin applying apparatus is illustrated in FIG. 3. 3A is a cross sectional view of the roll-on, which crosses two of the three balls. 3B is a cross sectional view through one of the three balls. 4 is a perspective view of an upper ball mounting portion (a) and a lower ball mounting portion (b), which are part of the roll-on.

Specifically, as can be seen in Figure 3, the cosmetic composition is stored in the lower container 10, the lower bowl mount 30 supports the lower portion of the ball 90, the upper bowl mount is fixed to the ball 90 Let's do it. The ball 90 is a spherical metal ball, and is rotated between the lower and upper ball mounting stand, the cosmetic composition is applied to the ball surface inside the container, and when the ball rotates, the applied cosmetic composition is transferred to the outside Applied to the skin. After use of the roll-on can be stored by closing the lid, the lid is the lid outer cap 50 and the inner cap 60 is overlapping. 3 is only an example of the present invention and is not limited thereto.

The cosmetic composition may further contain one or more of oat extract and yeast extract. At this time, the cosmetic composition is preferably contained in an arginine and lysine-containing polypeptide 0.01 to 2% by weight, oat extract 2 to 6% by weight and yeast extract 0.01 to 0.5% by weight based on the total weight of the composition, more preferably arginine And 0.01 to 1.5% by weight lysine containing polypeptide, 3 to 5% by weight oat extract and 0.06 to 0.2% by weight yeast extract. When contained in the content within the above range, while maintaining the effect of each active ingredient on the skin can maintain the skin stability.

In addition, the cosmetic composition may further contain one or more selected from the group consisting of adenosine, retinol and niacinamide. Preferably, the adenosine may be 0.01 to 0.07 wt% and the retinol may be 50 to 150 IU based on the total weight of the composition.

Arginine and lysine-containing polypeptides used in the present invention promote the synthesis of components of extracellular matrix such as collagen I, collagen III, collagen V, and decorin (proteoglycane) and inhibit MMP-1 release, thereby preventing aging. Has an effect. Typical of commercialized arginine and lysine containing polypeptides include PEPTISKIN® from Solabia ^. And the like. PEPTISKIN ^? Is a 40% dry extract solution composed of L-arginine and L-lysine oligopeptide, having a degree of polymerization of 2 to 7, and is obtained by a nonsolvent enzyme synthesis method.

Oat extract used in the present invention is obtained by extraction from oats (Oat). Oats Avena Also known as sativa , oats are rich in minerals such as protein, phosphorus, magnesium, and calcium, making them suitable for diet and effective for constipation. The oat extract used in the cosmetic composition of the present invention is extracted by a conventional grain extract manufacturing method. In one example, the ground product obtained by drying or grinding the oats may be extracted, for example, with a solvent selected from the group consisting of distilled water, organic solvents and mixtures thereof. In this case, the organic solvent used may be 1,3-butylene glycol, lower alcohol having 1 to 4 carbon atoms, ethyl acetate or acetone, and may be used two or more kinds sequentially according to the polarity of the organic solvent. Representative commercialized oat extracts include Silab's OSILIFT ^? And the like. OSILIFT ^? Oat kernel extract is an excellent material for immediate skin lifting and wrinkle relief.

The yeast extract used in the present invention is obtained by purifying and extracting yeast by microbial fermentation. Generally, yeast extract has a function of promoting the synthesis of collagen IV, collagen Ⅶ and Integrin, and has TGF beta1 activity. Therefore, it has an anti-wrinkle effect and a dermis-epidermis junction strengthening effect. Yeast extract is not limited as long as it is a general extraction method, it can be prepared by the following method. After dissolving the yeast (Saccharomyces Cerevisiae) in water, separating the dissolved and undissolved portions by filtration, the dissolved portions are taken separately, concentrated and filtered to obtain a yeast extract. Silab's TONISKIN ^? (Also called Yeast extract).

The skin care kit may contain the cosmetic composition inside the skin applicator, and may be included separately without the cosmetic composition contained in the skin applicator.

The present invention is a method of applying a cosmetic composition comprising the step of applying a tensile composition to the skin using a skin coating device after or simultaneously with the application of the cosmetic composition containing the arginine and lysine-containing polypeptide as an active ingredient to the skin To provide.

In the coating method, the skin coating apparatus may be a roll-on. The roll-on may have one, two or three or more balls.

Such roll-on is not limited as long as it is a form which makes the spherical roll contact skin.

The use of the skin applicator using the cosmetic composition according to the present invention is a cosmetic composition containing a yeast extract having a dermal-epidermal border strengthening effect and an oat extract having an instant wrinkle relieving effect through natural polys, and rolling; By the roll-on massage device having a massage effect and the effective ingredient penetration enhancement effect, it is possible to achieve the synergistic effect of skin cooling and elasticity enhancement.

In particular, when using the roll-on type skin coating device, there is no need to use a hand, the cosmetic composition is not absorbed by the hand can be applied to a larger amount of the composition to the application site, there is an advantage that can be used from time to time. In addition, there is no restriction on feeling such as stickiness or back feeling felt in the hand, and an excessive amount of a firming agent, a polymer, or the like can be used. In particular, the use of a roll-on type skin coating device provides a massage effect as the tension stimulus is applied to the skin, thereby improving blood flow, promoting the absorption of active ingredients, and improving skin cooling, wrinkle improvement and elasticity. There is this.

The present invention is described in more detail through the following implementation. However, the examples are for illustrating the present invention, and the scope of the present invention is not limited thereto.

[ Experimental Example  1] Expression level measurement experiment of collagen

The relative expression level of collagen was measured for skin fibroblasts. First, the comparison was made before and after giving tensile stimulation to dermal fibroblasts. Tensile stimulation was performed at 0.5 Hz and 17% of tension for 1 hour. Furthermore, arginine and lysine-containing polypeptide as a peptidomimetic skin ^(PEPTISKIN?, Manufactured by Solabia社) were compared after a given treatment, the tensile stimulation cycle before the tensile stimulation of the same conditions as in the skin fibroblasts. The results are shown in FIG.

As can be seen in Figure 2, it was confirmed that the collagen expression of the skin fibroblasts increased only by tensile stimulation. In addition, the highest level of collagen expression was observed when the peptiskin-treated dermal fibroblasts were subjected to tensile stimulation. In other words, when a tensile stimulus and peptiskin are given at the same time shows a synergistic effect and showed the best effect.

[ Example  One]

As the oat extract, an oat extract having the trade name Osiris (OSILIFT ^?, Manufactured by Silab) was used. As arginine and lysine-containing polypeptide was used as a trade name of a polypeptide is a peptidomimetic skin ^(PEPTISKIN?, Manufactured by Solabia社). Yeast extract was used as a yeast extract (manufactured by ^TONISKIN?, Silab社) Name Tony skin. 4% by weight of Osiris, 0.01% by weight of peptiskin, 0.5% by weight of toniskin, 0.04% by weight of adenosine and 100 IU of retinol were mixed. To add the material in the ratio as shown in Table 1 Essence was prepared.

The essence thus prepared was injected into a roll on applicator which is a skin applicator. The roll on applicator used is as shown in FIG. 3.

Input material Purified water To 100 Butylene Glycol 15.00 Adenosine 0.04 Niacin amide 0.10 Yeast extract (Toniskin ^? ) 0.50 Osilift ^? 4.00 Arginine / Lysine Polypeptide (Peptiskin ^? ) 0.01 Beta Glucan 0.50 Hydroxypropyl bispalmitamide M 0.01 Tromethine 0.05 P Polyoxyethylene Hydrogenated Castor Oil 0.30 Octyldodec-16 0.40 Carbomer 0.07 Xanthan gum 0.05 Disodium iodide 0.05 1,2 hexanediol 1.50 100

[ Experimental Example  2] skin Cooling  Experiment

With the essence prepared in Example 1, a clinical experiment was conducted on 10 subjects. The ten subjects were 32 ± 3.2 years old. The test was conducted with the right eye of each test subject as the test group and the left eye as the control group.

The test subjects were acclimated at 22 to 28 ° C. and relative humidity of 108 to 42% for 15 minutes in a constant temperature and humidity room after cleansing. The test group of the test subject was massaged for 30 seconds with a roll-on applicator, which is a skin coating apparatus according to the present invention, and the essence of Example 1 was applied, and the control group was absorbed by hand applying the same amount of fluid. Thereafter, the face was photographed before and immediately after application of the essence.

Skin temperature measurement cameras (IR camera, CAM PM575, FLIR. USA) were used to determine the immediate skin temperature change. The results are shown in FIGS. 5 to 7.

As can be seen in Figures 5 and 6, before and immediately after the use of the roll-on applicator, the skin temperature of the eyes was reduced, the skin temperature reduction rate was found to be much larger than the control. The temperature distribution photographs of the eye area skin of two test subjects photographed by a camera are shown in FIG. 7. Also from Figure 7, it can be seen that the difference in skin temperature reduction in the test group is more pronounced, demonstrating that the present invention is excellent in the instant cooling effect.

[ Experimental Example  3] skin wrinkle test

With the essence prepared in Example 1, the test subject and the test group and the control group were subjected to the skin wrinkle test in the same manner as in Experiment 2.

It was measured before, after application of the essence of Example 1, 2 weeks, 4 weeks and 8 weeks after use.

When photographing the face, the wrinkles around the eyes were photographed using a wrinkle measuring device (primos, GFMESSTECHNIK. The measurement result was shown to FIG. 8 and FIG. 9, and the photographic image is shown to FIG. Table 2 below shows a significant difference between paired t-test results before use, and Table 3 shows a significant difference between groups as independent t-test results.

Significance difference before use p-value after 2 weeks After 4 weeks 8 weeks later Test group 0.791 0.653 0.056 Control group 0.135 0.314 0.421

Difference between groups Week 0 after 2 weeks After 4 weeks 8 weeks later p-value 0.742 0.686 0.718 0.043

As can be seen in Figures 8 and 9, after 8 weeks of use of the test group product, the wrinkles around the eyes decreased by 11.7%. On the other hand, the control product did not show wrinkle reduction effect until 8 weeks. In addition, as can be seen in Figure 10, after 8 weeks of use compared to 0 weeks, it was possible to observe a decrease in the wrinkles around the eyes. In addition, as shown in Table 2 and Table 3, it was confirmed that after 8 weeks of use of the product, there was a significant difference (significant difference * p <0.05) between the groups and the group in the wrinkle improvement efficacy between the test group and the control group. This may be due to the promotion of absorption of the active ingredient by rolling the roll-on applicator.

[ Experimental Example  4] skin elasticity test

Having the essence prepared in Example 1, the test subject and the test group and the control group was subjected to the skin elasticity experiment in the same manner as in Experiment 2.

It was measured before, after application of the essence of Example 1, 2 weeks, 4 weeks and 8 weeks after use.

When the face was taken, the skin elasticity was measured using a skin elasticity measuring instrument (Cutometer 580 ^ⓡ , C + K, Germany). The results are shown in FIGS. 11 and 12.

As can be seen in Figures 11 and 12, after 2 weeks, 4 weeks and 8 weeks of using the test group product, it was found that the tendency of eyelid elasticity increased by 0.4%, 6.6%, and 9.1% (p = 0.062), respectively. . In contrast, the control product showed only 2.7% increase in eye elasticity by 8 weeks. This seems to be due to the eye massage effect by the roll-on applicator.

[ Experimental Example  5] Blood flow measurement experiment

With the essence prepared in Example 1, the test subject and the test group and the control group was measured in the same manner as in Experiment 2 to measure the blood flow of the skin. The results are shown in Figure 13 (a) blood flow change comparison graph according to the application method, b) blood flow image).

As can be seen in Figure 13a, the blood flow was increased by about 40% after use than before using the roll-on applicator of the present invention, the control using the hand rather the blood flow was reduced. In addition, as can be seen in Figure 13b, the value of the perfusion color scale is increased, which means that the speed of blood flow is faster toward the red color, and the value is lower, indicating that the speed of blood flow is slower toward the cold blue color, compared to before application It was confirmed that the speed of blood flow has increased. As a result, it was confirmed that the blood flow increased due to the massage effect by the roll-on applicator of the present invention.

[ Experimental Example  6] Skin absorption promoting effect experiment

With the essence prepared in Example 1, the test subject and the test group and the control group was measured in the same manner as in Experiment 2 to measure the degree of absorption of the skin of the active ingredient. The results are shown in Figure 14 (a) graph of absorption amount of the active ingredient according to the coating method, b) graph of absorption rate comparison).

As can be seen in Figure 14a, when using the roll-on applicator of the present invention the absorption of the active ingredient increased by about 40% compared to the case of hand coating, and as shown in Figure 14b, the absorption rate was increased about 2 times. As a result, when using the roll-on applicator of the present invention, it was confirmed that the skin delivery amount of the active ingredient is increased by the physical stimulation of the roll-on, and the initial absorption rate is increased by the massage effect by the roll-on.

10 lower container
20 upper container
30 Upper Ball Mount
40 Lower Ball Mount
50 lid outer cap
60 cap inner cap
70 lids decoration
80 liner
90 ball

Claims (16)

A cosmetic composition comprising an arginine and a lysine-containing polypeptide as an active ingredient and a skin applicator for applying the cosmetic composition to the skin. The method of claim 1,
The skin applicator is a skin care kit that is one or more selected from the group consisting of a roller, acupressure and a massage device to give a tensile stimulation. The method of claim 1,
The skin applicator is roll-on (roll-on) skin care kit. The method of claim 3,
The roll-on (roll-on) is a skin care kit that has three or more balls. The method of claim 1,
The cosmetic composition is a skin care kit that further contains one or more of oat extract and yeast extract. 6. The method of claim 5,
The cosmetic composition is a skin care kit containing 0.01 to 2% by weight of the arginine and lysine-containing polypeptide, 2 to 6% by weight oat extract and 0.01 to 0.5% by weight yeast extract based on the total weight of the composition. The method according to claim 1 or 5,
The cosmetic composition is a skin care kit that further contains one or more selected from the group consisting of adenosine, retinol and niacinamide. The method of claim 1,
The skin care kit is a skin care kit having one or more selected from the group consisting of skin cooling use, skin elasticity promotion use and skin wrinkle improvement use. A method of applying a cosmetic composition comprising a step of applying tensile stimulation to the skin using a skin applicator after applying or applying a cosmetic composition containing arginine and a lysine-containing polypeptide as an active ingredient to the skin. The method of claim 9,
The coating method is a method for applying a cosmetic composition having at least one selected from the group consisting of skin cooling use, skin elasticity use and skin wrinkle improvement. The method of claim 9,
The skin coating device is a method of applying a cosmetic composition is one or more selected from the group consisting of a roller, acupressure and a massage device to give a tensile stimulation. The method of claim 9,
The skin coating device is a roll-on (roll-on) is a coating method of the cosmetic composition. 13. The method of claim 12,
The roll-on is a coating method of the cosmetic composition is three or more balls. The method of claim 9,
The cosmetic composition is a skin care kit that further contains one or more of oat extract and yeast extract. The method according to claim 9 or 14,
The cosmetic composition is a method for applying a cosmetic composition further comprises one or more selected from the group consisting of adenosine, retinol and niacinamide. The method of claim 9,
The cosmetic composition is a method for applying a cosmetic composition containing 0.01 to 2% by weight of the arginine and lysine-containing polypeptide, 2 to 6% by weight oat extract and 0.01 to 0.5% by weight yeast extract.

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